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A probe amplification method based on multiple extension ligation, its use and kit

A technology for linking probes and capturing probes, which is applied in the field of detecting the SNP of the glucose-6-phosphate dehydrogenase gene, and can solve problems such as long detection time, incomplete enzyme digestion, and heavy workload

Active Publication Date: 2019-12-20
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

[0025] Other techniques, such as PCR restriction enzyme digestion, have high specificity, but are prone to incomplete digestion; HRM methods are highly sensitive, but can only detect one mutation site at a time, which is not suitable for detecting multiple allele mutation sites. The workload is heavy; DNA sequencing is the "gold standard" for mutation detection, but due to its long detection time and high cost, it is not suitable for clinical large-scale population screening

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  • A probe amplification method based on multiple extension ligation, its use and kit
  • A probe amplification method based on multiple extension ligation, its use and kit
  • A probe amplification method based on multiple extension ligation, its use and kit

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Embodiment Construction

[0052]In the following embodiments, the inventors disclosed 23 point mutations in the G6PD gene detected using the MELPA detection technology established in this study (see Table 1 above), and compared them with the iPLEX method and the direct sequencing method.

[0053] 1. The principle of MELPA technology and its application to detect 23 G6PD genotypes

[0054] Principle of MELPA technology

[0055] The principle of MELPA technology is to design a set of capture probe (capture probe, CP) and ligation probe (ligation probe, LP) near the detection gene target site (or SNP site), after lysing the blood sample or DBS sample, directly inject Genomic DNA is captured in the wells of the solid-phase plate, and then multiple probe extension and ligation reactions are performed. The analytes generated are tagged with universal primers, and then equivalent PCR amplification is performed by universal primers (universal primers, UP). The target fragment was obtained by increasing, and t...

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Abstract

The present invention relates to a probe amplification method based on multiplex extending connection, and uses thereof, and a kit, further to uses of the probe amplification method based on multiplex extending connection in detection of glucose-6-phosphate dehydrogenase gene SNP, and a kit for the method.

Description

technical field [0001] The present invention relates to the technical field of probe amplification method based on multiple extension ligation. More particularly, the present invention relates to the use of the method according to the present invention for detecting the SNP of the glucose-6-phosphate dehydrogenase gene; and the kit used in the method of the present invention. Background technique [0002] Glucose-6-phosphate dehydrogenase (G6PD) is an important enzyme in the cytoplasm, widely present in all cells, and especially important for the integrity and normal function of red blood cells. Glucose-6-phosphate dehydrogenase deficiency (G6PDd) is the most common hereditary enzyme disease. It is estimated that at least 400 million people in the world carry G6PD-deficient genes, of which about 350 million are distributed in malaria-endemic countries, see Nkhoma ET, Poole C, Vannappagari V, Hall SA, Beutler E: The globalprevalence of glucose-6- Phosphate dehydrogenase def...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858C12Q1/6883
Inventor 郑直田晓怡程志斌
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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